Figure 2: C11orf83 is a potent antiviral protein.
From: Mitochondrial C11orf83 is a potent Antiviral Protein Independent of interferon production
(A–E) Effects of C11orf83 overexpression to rVSV-GFP replication in HEK293 cells. All analysis was performed at 12 hours post rVSV-GFP (MOI = 1.0) infection. **p < 0.01. (A) Representative imaging fluorescent shows the replication of rVSV-GFP in cells. (B) Quantitative analysis of GFP positive cells by using flow cytometry. (C) Quantitative analysis of virus titer in cells supernatants. (D) Transcription detection of C11orf83 and N gene of VSV using RT-PCR. (E) Quantitative analysis of VSV-induced cells death using Trypan Blue staining. (F) Graphical representation of the Human genomic loci of C11orf83 showed the targeting sites for Cas9, sgRNA, and mutation initiation site. The sgRNA targeting sequences are labeled in magenta. The PAM sequences are labeled in red. The mutation initiation sites are indicated by the black triangle and the DNA scissors. (G) Detection of cells viability using MTT.(H–L) Effects of the loss of C11orf83 to rVSV-GFP replication in HEK293 cells. All analysis was performed at 12 hours post rVSV-GFP (MOI = 0.1) infection. Cas9-Control, HEK293 cells transfected with Cas9 and control sgRNA. Cas9-KO#1 and Cas9-KO#2, HEK293 cells transfected with Cas9 and sgRNA#1 or sgRNA#2 as shown in F. **p < 0.01. (H) Representative imaging fluorescent shows the replication of rVSV-GFP in cells. (I,K) Quantitative analysis of GFP positive cells and mean fluorescence intensity in cells by using flow cytometry. (J) Quantitative RT-PCR results shows the relative change of N gene transcription. (L) Quantitative analysis of VSV-induced cells death using Trypan Blue staining.
